Skin care compositions containing an organic extract of chick pea

ABSTRACT

Organic chick pea extracts are phytoestrogens, if present in an amount such as to provide an estrogenic activity equivalent to at least 1 nM of estradiol. Cosmetic compositions containing organic chick pea extracts are useful in improving the appearance of wrinkled, lined, dry, flaky, aged or photodamaged skin and improving skin thickness, elasticity, flexibility and plumpness.

This is a continuation of Ser. No. 08/901,052 filed Jul. 25, 1997, nowU.S. Pat. No. 6,030,620.

FIELD OF THE INVENTION

Cosmetic compositions containing an organic extract of chick peas andmethods of conditioning skin by applying such compositions to the skin.

BACKGROUND OF THE INVENTION

The human skin consists of two major layers, the bottom thicker layerdermis, and the top thinner layer—the epidermis. Dermis is the layerwhich provides the strength, elasticity and the thickness to the skin.

The main cell type of the dermis is fibroblasts, which is responsiblefor synthesis and secretion of all the dermal matrix components such ascollagen, elastin and glycosaminoglycans. Collagen provides thestrength, elastin the elasticity and glycosaminoglycans the moistnessand plumpness of the skin. With aging, the thickness of the dermal layeris reduced and this is believed to be partially responsible for theformation of wrinkles in aging skin. The top layer of human skin or theepidermis which provides the resilience and the barrier properties ofthe skin, is composed of many different cell types includingkeratinocytes, melanocytes and langerhans cells. Keratinocytes are themajor cell type of the epidermis (75-80% of the total number of cells inthe human epidermis). Richards et al. reported that estrogen stimulatessecretion of a protein, prolactin, by human dermal fibroblast cells andthat prolactin then stimulates proliferation of keratinocytes. Richardset al., Human Dermal Fibroblasts Express Prolactin In Vitro., J. Invest.Dermatol., 106: 1250, 1996.

Estrogens and synthetic compounds which act like estrogens are known toincrease the thickness of the dermal layer and reduce wrinkle formationin the aging skin. The changes in the skin such as skin dryness, loss ofskin elasticity and plumpness occurring after menopause is attributed tothe lack of estrogen production. Estrogen therapy prevents or slows downmany of these changes associated with aging skin (Creidi et al., Effectof a conjugated estrogen cream on aging facial skin, Maturitas, 19, p.211, 1994). Some of the effects of estrogen on skin include: increase inskin thickness and disappearance of fine wrinkles, increase of themitotic rate of the epidermis, reduction in the size and activity of thesebaceous gland, slow down of the rate of hair growth, stimulation ofcollagen turnover and increase in the production of hyaluronic acid andglycosaminoglycan synthesis of the fibroblasts (Pugliese, Menopausalskin, Skin Inc., March/April 1994: p 69-77).

In recent years, phytoestrogens (i.e., natural compounds which haveestrogen-like activity and which are found in plants) have beenincreasingly used for therapeutic purposes. Some of the uses describedare as hypocholesterolemic and antiatherogenic agents, treatment ofcardiovascular diseases especially in postmenopausal women, treatmentfor osteoporosis in the elderly and as an anticancer agent especiallyagainst breast cancer, endometrial and cervical cancer in women (Knightet al., Phytoestrogens—a short review, Maturitas, 22: 167-75, 1995).

The consumer demand for “natural” based products has been growing inrecent years. The consumers perceive chemical synthesis asenvironmentally unsafe. A chemically synthesized ingredient may containharsh chemicals. Natural products are perceived as pure and mild andsuperior to chemically synthesized products. However, delivering acosmetic benefit from plant sources is not trivial. In order to derive areal benefit from a “natural” source not only a plant or a part of theplant containing a specific active has to be identified, but a minimumconcentration and/or a specific extract of that plant has to beidentified which truly delivers a cosmetic benefit.

Over 500 compounds present in plants have been described to haveestrogenic activity. These compounds, collectively calledphytoestrogens, are found in a diverse number of plants includingcereals, legumes (including chick peas) and grasses (Price et al.,Naturally occurring estrogens in foods- a review., Food additives andcontaminants., 2, p. 73-106, 1985). Their concentrations vary in thedifferent parts of the plants, geographical locations, year of growthetc. Two major classes of plant compounds which possess phytoestrogenicactivity are flavonoids and coumestans. Some of the commonly describedphytoestrogenic compounds are genistein, biochanin A, formononetin,daidzein and their glycoside derivatives (Knight et al.,Phytoestrogens-a short review. Maturitas, J. Climactreic andpost-menopause, 22, p.167-75, 1995).

Chick pea or Spanish pea (Cicer arietinum), a common dietary lentil,contains flavonoids including daidzein, formononetin, biochanin A,pratensein, homoferreirin, medicarpin, maackiain, methyl coumestrol,medicagol, formononetin glucoside and biochanin A glucoside (Ingham etal., In: Progress in the chemistry of Organic natural products, vol 43:Ed-W. Herz et al., Springer-Verlag, Wien, N.Y., 1983). The flavonoidsfrom chick pea have been reported to have lipid lowering effects in theblood and liver of rats. Several nutritional studies report on theprotein from chick pea for use as nutritional supplements and ways toimprove the protein quality of chick pea. Vasiliou, U.S. Pat. No.4,761,285 discloses the use of chick peas as a dietary supplement or forinternal or topical treatment of hemorrhoids. In India, a cosmetic maskor skin treatment made from chick pea powder mixed with water is acommon beauty treatment.

The art discussed above does not describe organic chick pea extracts forskin care or cosmetic use. The art does not teach topical application oforganic chick pea extracts, as a phytoestrogen, and with a specifiedestrogenic activity.

SUMMARY OF THE INVENTION

The present invention includes a skin care composition comprising:

(i) an organic extract of chick peas in an amount such as to provide anestrogenic activity equivalent to at least 1 nM of estradiol, theestrogenic activity being determined by the test described herein;

(ii) a cosmetically acceptable vehicle.

The present invention also includes a method of improving or preventingthe condition of wrinkled, lined, dry, flaky, aged or photodamaged skinand improving skin thickness, elasticity, flexibility and plumpness,which method includes applying to the skin the inventive composition.Compositions of the invention are intended for topical application tomammalian skin which is already dry, flaky, lined, wrinkled, aged,photodamaged, or the inventive compositions may be appliedprophylactically to normal healthy skin to prevent or reduce thedeteriorative changes.

The present invention also includes a cosmetic method of increasingfibroblast and epidermal skin cell proliferation in human skin byapplying to the skin the inventive composition.

DETAILED DESCRIPTION OF THE INVENTION

Except in the examples, or where otherwise explicitly indicated, allnumbers in this description indicating amounts of material or conditionsof reaction, physical properties of materials and/or use are to beunderstood as modified by the word “about.” All amounts are by weight ofthe composition, unless otherwise specified.

Chick Peas

Chick peas are suitable for use in the inventive compositions in theform of an organic extract. The chick pea extract is prepared for use inthe present invention from dried chick peas. Dried chick peas may beobtained from Arrowhead Mills, from health food stores or supermarkets.

The organic chick pea extracts are prepared by extracting the. driedchick peas with a solvent by stirring 1 part of dried chick peas with 2to 5 parts of the solvent for from 4 to 24 hours at room temperature.Suitable solvents are described hereinbelow. The extracts are clarifiedby filtration and/or centrifugation, then dried by evaporation(optionally, under vacuum) to obtain the organic chick pea extract.

Solvents suitable for: the preparation of chick pea extract for useherein include, but are not limited to: ethanol, methanol, hexane,chloroform, dichloromethane and ethyl acetate. The preferred solventsare dichloromethane, methanol, or ethanol in order to optimize activity.The extract may be further concentrated, fractioned, re-extracted orpurified, e.g. by organic solvent extraction or by chromatography.

It has been found, as part of the present invention, that the organicchick pea extract has to be employed in a specific minimum amount todeliver an estrogenic activity.

Chick pea extract or powder is employed in an amount of from 0.0001 to50%, as long as it delivers the estrogenic activity equivalent to atleast 1 nM, generally in the range of from 1 nM to 100 nM, of estradiol.Estrogenic activity of the chick pea extract is determined by comparisonto the activity of 1 nM estradiol using ZR-75 cells in a side by sideexperiment, as described in Example 1 below. Preferably, the chick peaextract is employed in an amount such as to deliver the estrogenicactivity equivalent to at least 2 nM of estradiol.

Cosmetically Acceptable Vehicle

The composition according to the invention also comprises a cosmeticallyacceptable vehicle to act as a diluant, dispersant or carrier for thechick pea extract in the composition, so as to facilitate itsdistribution when the composition is applied to the skin.

Vehicles other than or in addition to water can include liquid or solidemollients, solvents, humectants, thickeners and powders. An especiallypreferred nonaqueous carrier is a polydimethyl siloxane and/or apolydimethyl phenyl siloxane. Silicones of this invention may be thosewith viscosities ranging anywhere from about 10 to 10,000,000mm^(2/)s(centistokes) at 250° C. Especially desirable are mixtures oflow and high viscosity silicones. These silicones are available from theGeneral Electric Company under trademarks Vicasil, SE and SF and fromthe Dow Corning Company under the 200 and 550 Series. Amounts ofsilicone which can be utilized in the compositions of this inventionrange anywhere from 5% to 95%, preferably from 25% to 90% by weight ofthe composition.

The cosmetically acceptable vehicle will usually form from 5% to 99.9%,preferably from 25% to 80% by weight of the composition, and can, in theabsence of other cosmetic adjuncts, form the balance of the composition.Preferably, the vehicle is at least 80 wt. % water, by weight of thevehicle. Preferably, water comprises at least 50 wt. % of the inventivecomposition, most preferably from 60 to 80 wt. %, by weight of thecomposition.

Optional Skin Benefit Materials and Cosmetic Adjuncts

An oil or oily material may be present, together with an emulsifier toprovide either a water-in-oil emulsion or an oil-in-water emulsion,depending largely on the average hydrophilic-lipophilic balance (HLB) ofthe emulsifier employed.

The inventive compositions preferably include sunscreens. Sunscreensinclude those materials commonly employed to block ultraviolet light.Illustrative compounds are the derivatives of PABA, cinnamate andsalicylate. For example, octyl methoxycinnamate and 2-hydroxy-4-methoxybenzophenone (also known as oxybenzone) can be used. Octylmethoxycinnamate and 2-hydroxy-4-methoxy benzophenone are commerciallyavailable under the trademarks, Parsol MCX and Benzophenone-3,respectively. The exact amount of sunscreen employed in the emulsionscan vary depending upon the degree of protection desired from the sun'sUV radiation.

Emollients are often incorporated into cosmetic compositions of thepresent invention. Levels of such emollients may range from 0.5% to 50%,preferably between 5% and 30% by weight of the total composition.Emollients may be classified under such general chemical categories asesters, fatty acids and alcohols, polyols and hydrocarbons.

Esters may be mono- or di-esters. Acceptable examples of fatty di-estersinclude dibutyl adipate, diethyl sebacate, diisopropyl dimerate, anddioctyl succinate. Acceptable branched chain fatty esters include2-ethyl-hexyl myristate, isopropyl stearate and isostearyl palmitate.Acceptable tribasic acid. esters include triisopropyl trilinoleate andtrilauryl citrate. Acceptable straight chain fatty esters include laurylpalmitate, myristyl lactate, and stearyl oleate. Preferred estersinclude coco-caprylate/caprate (a blend of coco-caprylate andcoco-caprate), propylene glycol myristyl ether acetate, diisopropyladipate and cetyl octanoate.

Suitable fatty alcohols and acids include those compounds having from 10to 20 carbon atoms. Especially preferred are such compounds such ascetyl, myristyl, palmitic and stearyl alcohols and acids.

Among the polyols which may serve as emollients are linear and branchedchain alkyl polyhydroxyl compounds. For example, propylene glycol,sorbitol and glycerin are preferred. Also useful may be polymericpolyols such as poly-propylene glycol and polyethylene glycol. Butyleneand propylene glycol are also especially preferred as penetrationenhancers.

Exemplary hydrocarbons which may serve as emollients are those havinghydrocarbon chains anywhere from 12 to 30 carbon atoms. Specificexamples include mineral oil, petroleum jelly, squalene andisoparaffins.

Another category of functional ingredients within the cosmeticcompositions of the present invention are thickeners. A thickener willusually be present in amounts anywhere from 0.1 to 20% by weight,preferably from about 0.5% to 10% by weight of the composition.Exemplary thickeners are cross-linked polyacrylate materials availableunder the trademark Carbopol from the B.F. Goodrich Company. Gums may beemployed such as xanthan, carrageenan, gelatin, karaya, pectin andlocust beans gum. Under certain circumstances the thickening functionmay be accomplished by a material also serving as a silicone oremollient. For instance, silicone gums in excess of 10 centistokes andesters such as glycerol stearate have dual functionality.

Powders may be incorporated into the cosmetic composition of theinvention. These powders include chalk, talc, kaolin, starch, smectiteclays, chemically modified magnesium aluminum silicate, organicallymodified montmorillonite clay, hydrated aluminum silicate, fumed silica,aluminum starch octenyl succinate and mixtures thereof.

Other adjunct minor components may also be incorporated into thecosmetic compositions. These ingredients may include coloring agents,opacifiers and perfumes. Amounts of these other adjunct minor componentsmay range anywhere from 0.001% up to 20% by weight of the composition.

Use of the Composition

The composition according to the invention is intended primarily as aproduct for topical application to human skin, especially as an agentfor conditioning, moisturizing and smoothening the skin, increasing itsthickness, flexibility and elasticity and preventing or reducing theappearance of wrinkled, lined or aged skin.

In use, a small quantity of the composition, for example from 1 to 100ml, is applied to exposed areas of the skin, from a suitable containeror applicator and, if necessary, it is then. spread over and/or rubbedinto the skin using the hand or fingers or a suitable device.

Product Form and Packagina

The topical skin care composition of the invention can be formulated asa lotion, a cream or a gel. The composition can be packaged in asuitable container to suit its viscosity and intended use by theconsumer. For example, a lotion or a cream can be packaged in a bottleor a roll-ball applicator, or a propellant-driven aerosol device or acontainer fitted with a pump suitable for finger operation. When thecomposition is a cream, it can simply be stored in a non-deformablebottle or squeeze container, such as a tube or a lidded jar. Thecomposition may also be included in capsules such as those described inU.S. Pat. No. 5,063,507, incorporated by reference herein. The inventionaccordingly also provides a closed container containing a cosmeticallyacceptable composition as herein defined.

The following specific examples further illustrate the invention, butthe invention is not limited thereto. All p-values in the Examples werecalculated using a student t-test.

EXAMPLE 1

This example illustrates that the organic chick pea extract at aconcentration such as to provide a phytoestrogenic activity equivalentto at least 1 nM of estradiol is suitable for use in the presentinvention.

Preparation of Chick Pea Extracts:

Dried chick peas were obtained from Arrowhead Mills (Hareford, Tex.) andground to a powder in a dry grinder. The chick pea powder thus obtainedwas extracted using dichloromethane (DCM) or methanol (1:3 w/v) bymixing the dry powder with the solvent for 20 hours at room temperature.The extracts were clarified by filtration and centrifugation. Theextracts were then dried by evaporation under vacuum to obtain the dryextract. Extraction of 63 g of chick pea dry powder with DCM dissolved1.47 g of dry material; extraction of 60 g of chick pea powder withmethanol dissolved 2.434 g of dry material. This amounts to extractionof 2.31% by DCM and 4.06% by methanol. This dry extract was thenredissolved in dimethyl sulfoxide (DMSO) at 50 μg/μl concentration fordosing the cells.

Methodology Used for Determining the Rate of DNA Synthesis in Cells:

The incorporation of ³H-thymidine by cultured cells was used as an assayof cell proliferation (ZR75 or keratinocyte). Thymidine is one of fourdeoxynucleosides which are the monomeric units of DNA. Prior to celldivision of a somatic cell, the complete genome of the cell undergoingcell division is replicated. This involves large scale DNA synthesis bythe cell and enables both daughter cells to receive identical copies ofthe genetic material. When ³H-thymidine is included in the culture mediaof cells which are synthesizing DNA in preparation for cell divisionthen the labeled thymidine is incorporated into the newly synthesizedDNA. The extent of incorporation of ³H-thymidine into a population ofcells is proportional to the rate of DNA synthesis. by this populationof cells and therefore an indication of their cellular proliferation.

The following test was employed to determine whether organic chick peaextracts have phytoestrogenic activity -and whether this activity isequivalent to at least 1 nM estradiol:

ZR75 cell line is a ductal breast carcinoma cell line, originallyisolated from malignant mammary epithelium of a sixty-three year oldCaucasian female (Engel et al., Human breast carcinoma cells incontinuous culture: A review., Cancer Res., 38: 4327-4339, 1978) Thiscell line contains receptors for estrogen, progesterone and othersteroid hormones, but responds through an increase in proliferation onlyto estrogen. The cell line contains high affinity estrogen-specificreceptors. Therefore, this cell line is used for testing estrogen-likeactivity (Markiewicz et al., In vitro bioassays of non-steroidalphytoestrogens, J. Steroid Biochem. Molec. Biol., 45: 399-405, 1993).

1. ZR75 cells (from American Type Culture Collection, Rockville, Md.)were grown in RPMT1640 media (from Gibco Life Technologies) with 10%fetal bovine serum (FBS), 100 units penicillin per ml and 100 units ofstreptomycin per ml. The media did not contain Phenol Red (a weakestogen mimetic). The. cells were seeded at a density of one million per75 cm2 flask. For the experiment, the cells were seeded in 24 wellplates at 100,000 cells per ml per well.

2. After growing for 24 hours, the media was removed, the cells werewashed with PBS and 1 ml of RPMI 1640 without serum (but withstreptomycin and penicillin) was added. Stock solutions of chick peaextract in dimethyl sulfoxide. (DMSO) and estradiol in water wereprepared. Various concentrations of organic chick pea extract andestradiol, as indicated in Table 1, were then dosed directly into eachwell. After another 24 hours, one loci of [methyl-3H] thymidine wasadded to each well. The media was removed after 24 hours. The cells werewashed once in PBS, the PBS was removed completely and the cells wereleft on ice to incubate with 1 ml per well of10% TCA (trichlioroaceticacid) for 30 minutes. The plates were washed 3 times with 5% TCA toremove all traces of thymidine which wasn't incorporated into the cells.500 μl of 0.1M sodium hydroxide was added to each well and the plateswere incubated at room temperature for at least 30 minutes. 250 μl ofeach sample was transferred to scintillation vials and after adding 5 mlof counting fluid, the vials were counted for 5 minutes each on asetting for tritium. Data from triplicate wells were calculated as %thymidine incorporation into DNA compared to that of control wells whichdid not receive any chick pea extract or estradiol. Values wereexpressed as mean of triplicate wells ± standard deviation.

The results that were obtained are summarized in Table 1.

TABLE 1 ZR 75 DNA Chick pea DCM ZR 75 cell DNA synthesis extractsynthesis Estradiol (nM) (% of Control) (μg/ml) (% of Control) 0 100 ±10 0 100 ± 17.5 0.01 117 ± 19.7 0.01 106 ± 12.8 0.1 106 ± 16.3 0.1 176 ±25.6* 1.0 162 ± 3.2* 1.0 174 ± 16.6* 10.0 267 ± 15.1* 10.0 226 ± 28.7*100 203 ± 51.2* 1000 178 ± 20.1* *indicates statistically significantstimulation of DNA synthesis compared to control at p level < 0.05.

As can be seen from the results in Table 1, 1 nM (0.274 ng/ml) or higherconcentrations of estradiol increased DNA synthesis of ZR75 cells.Similar activity of this particular extract of chick peas was obtainedat a concentration of 0.1 μg/ml or higher.

The organic chick pea extract is suitable for use in the inventivecompositions in an amount such as to deliver estrogenic activityequivalent to at least 1 nM of estradiol. For this particular extract,concentration of 0.1 μg of chick peas per 1 ml of media delivered therequisite activity.

EXAMPLE 2

Example 1 was repeated twice with various concentrations of chick peaextract as indicated in Table 2.

The results that were obtained are summarized in Table 2.

TABLE 2 Treatment cpm ± SD % of Control p value Experiment 2: Control14566 ± 719  100 ± 4.9 Estradiol 122254 ± 3940  839 ± 27 0.000003 (10nM) Chick pea DCM 22530 ± 1302 154.6 ± 8.9  0.0016 extract (10 μg/ml)Experiment 3: Control 14302 ± 1669   100 ± 11.6 Estradiol 30158 ± 1087 211 ± 7.6 0.0004 (10 nM) Chick Pea DCM 26053 ± 901   182 ± 6.3 0.0009extract (1.0 μg/ml) Chick Pea DCM 49541 ± 591   346 ± 2.1 0.000005extract (10.0 μg/ml)

It can be seen from the results in Table 1 (representing Experiment 1)and Table 2 that in 3 separate experiments performed on 3 separate daysusing 3 different passages of the ZR75 cells, the organic chick peaextract stimulated ZR75 cell proliferation significantly.

EXAMPLE 3

This example illustrates that keratinocytes treated with an organicchick pea extract excreted a substance which stimulated theproliferation of fibroblasts.

Cell Culture of Keratinocytes:

Human keratinocytes, isolated from neonatal foreskin by trypsintreatment were grown in Dulbecco Modification Eagle (DME) Hams F12 (3:1)medium/5% fetal calf serum in the presence of mitomycin C treated 3T3mouse fibroblasts for establishing dividing keratinocyte colonies. Cellswere grown under the above condition until their second passage and keptfrozen for future use. Frozen second passage keratinocytes were thawedand plated into the above medium and grown for five days. On day 5, whenthe cells were 70-80% confluent, they were trypsinized and plated at20,000-30,000 cells per well in 24 well plates in a serum freekeratinocyte growth medium (KGX-from Clonetics, San Diego Calif.)containing 0.15 mM calcium. The cells were grown to 80% confluence for 5days before the experiment.

Cell Culture of Fibroblasts:

Neonatal human foreskin fibroblasts obtained from Clonetics Corporation(San Diego, Calif.) were cultured in DMEM containing 10% fetal bovineserum (both obtained from Life Technologies, Grand Island, N.Y.). Theexperiments were carried out in cells in their 5 to 15th passage. Cellswere plated at 7,000-10,000 cells per well in 24-well plates and grownto 80% confluence for 5 days. before the experiment.

Preparation of Keratinocyte Conditioned Medium:

For experiments looking for the interaction of keratinocytes andfibroblasts in response to treatment with chick pea extracts,keratinocyte conditioned medium was prepared as follows: 80% confluentkeratinocytes prepared as described before, were treated with variousconcentrations of chick pea extracts (as indicated at Table 3) for 24hrs in RPMI medium without serum and phenol red. The medium collectedfrom the plates is referred to as “keratnocyte conditioned medium.

Test Methodology:

Keratinocytes and fibroblasts were separately dosed with variousconcentrations of the chick pea extract in 1 ml of RPMI medium withoutserum and phenol red. Fibroblasts were also dosed with 1 ml ofkeratinocyte conditioned medium. After 24 hours, ³H thymidine was addedat 1 μCi per well and the cells were incubated for 24 hrs. Amount of ³Hthymidine associated with the cellular DNA were assessed as described instep 2 of Example 1.

The results that were obtained are summarized in Table 3.

TABLE 3 Keratinocyte conditioned medium on Keratinocytes Fibroblastsfibroblasts cpm ± SD cpm ± SD cpm ± SD Treatment (% of control) (% ofcontrol) (% of control) Control 105514 ± 19070 32206 ± 1207 24385 ± 3056(100 ± 18.1) (100 ± 3.7) (100 ± 12.5) Chick Pea DCM extract: 0.1 μg/ml109855 ± 28739 39617 ± 6046 47799 ± 3920 (104 ± 27.2) (123 ± 18.7) (196± 16.0) p = 0.838 p = 0.1057 p = 0.00132 1.0 μg/ml 105596 ± 19888 35237± 3279 42093 ± 5448 (100 ± 18.8) (109 ± 10.2) (172 ± 1.83) p = 0.996 p =0.2074 p = .0079

The results in Table 3 demonstrate that chick pea extracts F did nothave a direct effect on either keratinocyte or fibroblast proliferation.However, the chick pea extract induced keratinocytes to secrete growthpromoting substances for fibroblasts since conditioned medium fromkeratinocytes exposed to chick pea extract significantly increased theproliferation of fibroblasts.

EXAMPLE 4

This example illustrates that organic chick pea extracts increaseproliferation of cells in the epidermal portion of the piglet skin. Inthis example, estradiol, pure phytoestrogenic compounds, and chick peaextracts were tested for their effects on epidermal cell proliferation.

Piglet Skin Organ culture:

Pig skin is morphologically and biochemically similar to human skin andis therefore used commonly for testing the effects of materials for,human topical use. Fresh shaved skin from piglets was obtained from thelocal slaughter house, washed with Dove® Soap and dermatomed at 200μthickness to obtain the top epidermis and dermal layer. Uniform punchbiopsies of the dermatomed pig skin was taken using a 7 mm punch. Thebiopsies were washed in medium containing high antibiotic/antimycoticmix(Kanamycin and penicillin and streptomycin from Gibco) and incubatedfor 3 days in high glucose DMEM medium containing hydrocortisone,L-glutamine, antibiotic and antimycotic in the absence of serum intranswell plates, 3 biopsies per well. The medium was fed from thebottom of the transwell so that the epidermal side of the biopsies wasin contact with air. On the 3rd day, the medium was changed to freshmedium, and the biopsies were exposed to the various concentrations ofvarious actives in 1 μl of DMSO, as indicated in Table 4. 3 days later,medium was changed, actives readded and the biopsies were labeled with10 μCi of 3H thymidine per well. After washing the biopsies in PBS, theepidermis was split from the dermis by incubating the biopsies for 24hrs in 2 M sodium bromide solution with shaking. The epidermis waspeeled off from the dermis using a tweezer and dissolved in1 ml of 0.5NNaOH with overnight incubation at 60 ° C. 200 μl of the dissolvedepidermis was used for scintillation counting to determine the amount of³H thymidine incorporation into the epidermis. The data was calculatedas mean cpm for at least 4 replicates, and was also calculated as % ofcontrol biopsies which received no actives. The results that wereobtained are summarized in Table 4.

TABLE 4 Treatment cpm ± SD % of Control p value Experiment 1 Control4563 ± 1523  100 ± 33.3 Estradiol 1 nM 2690 ± 1113 58.9 ± 24.4 10 nM5190 ± 1096 113 ± 24  0.476 100 nM 1757 ± 1492 39.0 ± 32.6 IsoflavonoidsControl 3789 ± 1164  100 ± 30.7 Daidzein 1 μg/ml 8599 ± 927   227 ± 24.40.000226 10 μg/ml 11065 ± 2901   292 ± 76.5 0.00228 Formononetin 4770 ±1260  126 ± 33.2 0.826 1 μg/ml 10 μg/ml 6608 ± 2886  174 ± 76.1 0.1867Chick Pea DCM extract 0.1 μg/ml 10922 ± 3887  288 ± 102 0.0099 1.0 μg/ml13149 ± 3037  347 ± 80  0.000686 Experiment 2 Control 1155 ± 69   100 ±5.9  Estradiol 1 nM 2273 ± 783   196 ± 67.7 0.1147 Estradiol 10 nM 3817± 1175 330 ± 101 0.0292 Estradiol 100 nM 2785 ± 751  241 ± 65  0.0339Chick Pea Methanol extract Control 4451 ± 1961 100 ± 44  0.01 μg/ml 5503± 3026  123 ± 67.9 0.5463 0.1 μg/ml 8435 ± 2034  189 ± 45.6 0.0205 1μg/ml 2256 ± 704  50.6 ± 15.8 —

It can be seen from the results in Table 4 that in 2 separateexperiments, estradiol at 10 nM increased proliferation of the pigletepidermal cells (in experiment 2, the increase was statisticallysignificant) Daidzein also demonstrated this effect, daidzein being moreeffective than the natural hormone estradiol. Another isoflavone,formononetin had no significant growth stimulating effects on pigepidermis. Both methanolic and dichloromethane extracts of chick peaalso had growth promoting effect dichloromethane extract being moreeffective than methanolic extract. Dichlomethane extract of chick peawas even more effective than the pure isoflavones or estiradiol. Theresults demonstrate the beneficial effects of organic chick pea extracton skin cells in a model system which resembles very closely human skin.

Examples 5-10 illustrate skin care compositions according to the presentinvention. The” compositions can be processed in conventional manner.They are suitable for cosmetic use. In particular, the compositions aresuitable for application to wrinkled, lined, rough, dry, flaky, agedand/or UV-damaged skin to improve the appearance and the feel thereof aswell as for application to healthy skin to prevent or retarddeterioration thereof.

EXAMPLE 5

This example illustrates a high internal phase water-in-oil emulsionincorporating the inventive composition.

% w/w CHICK PEA DCM EXTRACT 0.5 1,3-dimethyl-2-imidazolidinone 0.2 Brij92* 5 Bentone 38 0.5 MgSO₄7H₂O 0.3 Butylated hydroxy toluene 0.01Perfume qs Water to 100 *Brij 92 is polyoxyethylene (2) oleyl ether

EXAMPLE 6

This example illustrates an oil-in-water cream incorporating theinventive composition.

% w/w CHICK PEA METHANOL EXTRACT 2 Mineral oil 41,3-dimethyl-2-imidazolidinone 1 Brij 56* 4 Alfol 16RD* 4Triethanolamine 0.75 Butane-1,3-diol 3 Xanthan gum 0.3 Perfume qsButylated hydroxy toluene 0.01 Water to 100 *Brij 56 is cetyl alcoholPOE (10) Alfol 16RD is cetyl alcohol

EXAMPLE 7

This example illustrates an alcoholic lotion incorporating thecomposition according to the invention.

% w/w CHICK PEA METHANOL EXTRACT 5 1,3-dimethyl-2-imidazolidinone 0.1Ethanol 40 Perfume qs Butylated hydroxy toluene 0.01 Water to 100

EXAMPLE 8

This example illustrates another alcoholic lotion containing theinventive composition.

% w/w CHICK PEA DCM EXTRACT 10 1,3-dimethyl-2-imidazolidinone 0.01Ethanol 40 Antioxidant 0.1 Perfume qs Water to 100

EXAMPLE 9

This example illustrates a suncare cream incorporating the compositionof the invention:

% w/w CHICK PEA ETHANOL EXTRACT 2 1,3-dimethyl-2-imidazolidinone 0.2Silicone oil 200 cts 7.5 Glycerylmonostearate 3 Cetosteryl alcohol 1.6Polyoxyethylene-(20)-cetyl 1.4 alcohol Xanthan gum 0.5 Parsol 1789 1.5Octyl methoxycinnate (PARSOL MCX) 7 Perfume qs Color qs Water to 100

EXAMPLE 10

This example illustrates a non-aqueous skin care compositionincorporating the inventive combination.

% w/w CHICK PEA DCM EXTRACT 5 1,3-dimethyl-2-imidazolidinone 1 Siliconegum SE-30¹ 10 Silicone fluid 345² 20 Silicone fluid 344³ 50.26 Squalene10 Linoleic acid 0.01 Cholesterol 0.03 2-hydroxy-n-octanoic acid 0.7Vitamin E linoleate 0.5 Herbal oil 0.5 Ethanol 2 ¹A dimethyl siliconepolymer having a molecular weight of at least 50,000 and a viscosity ofat least 10,000 centistokes at 25° C., available from GEC ²Dimethylsiloxane cyclic pentamer, available from Dow Corning Corp. ³Dimethylsiloxane tetramer, available from Dow Corning Corp.

It should be understood that the specific forms of the invention hereinillustrated and described are intended to be representative only.Changes, including but not limited to those suggested in thisspecification, may be made in the illustrated embodiments withoutdeparting from the clear teachings of the disclosure. Accordingly,reference should be made to the allowing appended claims in determiningthe full scope of the invention.

What is claimed is:
 1. A skin care composition comprising: (i) anorganic extract of chick peas in an amount such as to provide anestrogenic activity equivalent to at least 1 nM of estradiol. (ii) acosmetically acceptable vehicle.